ExtData_Figure_7_qPCR

(B) Testing of luciferase reporters with sequence insertions in the introns, as shown in diagram in A. HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing the 99 bp insertions in STOML2 or YKT6 either in an exon or an intron, with or without PR8 PA-X. RNA was extracted and luciferase mRNA levels were quantified by qRT-PCR and normalized to 18S. Two sets of luciferase primers were used - either both annealing to exons or one to exon and one to intron, to measure levels of the processed mRNA and unspliced pre-mRNA, respectively.